HIV (Human immunodeficiency virus) infection disease progression molecule marker miR-99a
A technology of mir-99a and molecular markers, applied in the field of miR-99a, a molecular marker of HIV infection disease progression, can solve the problems of lack of research and achieve the effect of simple operation, high sensitivity, and easy mass screening
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[0018] 1. Extraction of total RNA from peripheral blood mononuclear cells
[0019] 5ml of blood was drawn from 11 patients with rapid progress of HIV infection after 120 days of infection, and 5ml of blood from each of 6 typical progressors of HIV infection and 4 normal controls were taken as controls. Suspend the cells and place them in a 1.5ml RNase / DNase-free centrifuge tube.
[0020] Total RNA was extracted from peripheral blood mononuclear cells using an RNA extraction kit (Beijing Kuangbo Biotechnology Co., Ltd.), and 1 μl (20 nM) of the sequence 5'-CAACCUCCUAGAAAGA-3' (SEQ ID NO: 2) External Control-1 (synthesized by Shanghai Sangon Bioengineering Technology Co., Ltd.) to monitor the quality of RNA extraction from peripheral blood mononuclear cells. The concentration of extracted total RNA was measured using Thermo NanoDrop2000c.
[0021] 2. Three-step method for quantitative detection of miRNA in peripheral blood mononuclear cells
[0022] (1) Add polyA tail:
[00...
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